A simple and accurate ion chromatographic method was developed for the determination of sulfate content in presence of glutaric acid in (1S, 4R)-4- [2- amino- 6- (cyclopropyl amino) - 9 H- purin- 9-yl]-2- cyclopentene- 1-methanol hemisulfate, commercially known as Abacavir hemisulfate drug substance. In the titration method for sulfate content using alkaline solution, there is a high probability of interference of glutaric acid present at trace levels. Chromatographic separation between sulfate anion and glutarate anion has been achieved on Metrosep A supp-5 Ion exchange column using a mobile phase system containing a mixture of 1.0 mM sodium hydrogen carbonate, 6.0 mM sodium carbonate prepared in HPLC grade water, mixed with Acetonitrile in the ratio of 7:3 (v/v). The resolution between the two anions was found to be more than 5. The limit of detection (LOD) and limit of quantification (LOQ) of glutarate anion was 16 and 50 ng.mL-1 respectively, for 20 µL injection volume. The percentage recovery of sulfate ranged from 99 to 102 % w/w in the (1S, 4R)-4- [2- amino- 6- (cyclopropyl amino) - 9 H- purin- 9-yl]-2- cyclopentene- 1-methanol hemisulfate sample. The test solution and mobile phase were observed to be stable up to 48 h after preparation. The validated method produced good results of precision, linearity, accuracy, robustness and ruggedness. The proposed method was found to be suitable and accurate for the quantitative determination of sulfate in the bulk samples of Abacavir hemisulfate.